The present invention relates to a method of increasing the length of a telomere in an animal cell. The invention is practiced by administering an aqueous extract of an Uncaria species to an animal.
A telomere is a region of repetitive DNA sequence at the end of a chromosome, which protects the end of the chromosome from truncation and from fusion with a neighboring chromosome. Accordingly, telomeres provide stability to linear DNA molecules (Greider, C W (1990) BioEssays 12(8):363-369). The telomeric TTAGGG repeat sequences capping the ends of chromosomes have been shown to shorten during replicative ageing of normal cells (Harley et al. (1990) Nature 345(6274): 458-460).
Telomeres are consumed during cell division, but are replenished by the action of the enzyme telomerase reverse transcriptase. Human somatic cells without telomerase gradually lose telomeric sequences as a result of incomplete replication. Various methods for increasing the length of a telomere in a cell are known in the art (Joeng et al. (2004) Long lifespan in worms with long telomeric DNA. Nature Genetics 36(6): 607-611; Bodnar et al. (1998) Extension of life-span by introduction of telomerase into normal human cells. Science 279(5349): 349-352). These methods primarily rely on increasing the activity of telomerase. For example, a study that looked at normal human cells over-expressing the human telomerase catalytic subunit demonstrated elongated telomeres as a result of enhanced telomerase activity (Bodnar et al., supra) As distinguished from the present invention, these prior art methods do not provide a means of enhancing DNA repair in combination with a method of increasing the activity of telomerase, or otherwise increasing telomere length.
Methods for measuring telomere length are well known in the art and are described, for example, in Cawthon, R M (2002) Nucleic Acids Res. 30(10): e47, Harley et al. (1990) Nature 345: 458-460, O'Callaghan and Fenech (2011) Biological Procedures Online 13:3, U.S. Pat. No. 5,489,508 (West et al., issued Feb. 6, 1996), and U.S. Pat. No. 5,741,677 (Kozlowski et al., issued Apr. 21, 1998). Any method of measuring telomere length can be used in the present invention.
Any nucleated cell type may be used for the purpose of measuring telomere length. Lymphocytes and buccal cells are commonly used, as they are easily obtained from patients (O'Callaghan and Fenech (2011) Biological Procedures Online 13:3). Peripheral leukocytes are also commonly used for telomere length assessment (Aviv et al. (2011) Nucleic Acids Research 39(20): e134). As described herein, the terms “peripheral leukocytes” and “white blood cells” are used interchangeably. Genomic DNA isolated from the nucleated cell type of interest is used to measure telomere length. Methods for isolating genomic DNA are well known in the art.
AC-11°, formerly known as C-Med 100, is a hot water extract of the bark of the plant Uncaria tomentosa. Its characterization and method of preparation are described in U.S. Pat. No. 6,039,949 to Pero (issued Mar. 21, 2000). Animal and human studies have demonstrated a beneficial effect of AC-11® on enhancing DNA repair and immune function (Sheng et al. (2000) J. Ethnopharmacol. 69: 115-126; Sheng et al. (2001) Phytomedicine 8(4): 275-282).
U.S. Pat. No. 6,039,949 to Pero discloses a water soluble extract of an Uncaria species, as well as a method for preparing it. The reference further discloses oral administration of the extract to rats. The reference does not, however, contemplate a method of lengthening the telomeres or otherwise manipulating or maintaining telomere length. Likewise, the reference does not contemplate measuring telomere length.
Sheng et al. (2001, supra) disclose oral administration of C-MED-100 (a water extract of Uncaria tomentosa) to twelve healthy adult human volunteers. The doses tested were daily doses of 250 mg and 350 mg, for 8 weeks. A decrease of DNA damage and an increase in DNA repair was seen in both supplement groups when compared to an untreated control group. The reference does not disclose a method of lengthening telomeres and does not contemplate measuring telomere length before and after administration of the supplement.
Lamm et al. (2001, Phytomedicine 8(4):267-274) disclose oral administration of C-MED-100® at a dose of 350 mg×2 times daily for two months, to human volunteers. Study participants supplemented with C-MED-100® were tested for their ability to respond to a 23 valent pneumococcal vaccine as compared to an untreated control group. Statistically significant immune enhancement was observed for those in the supplemented group. The reference does not disclose a method of lengthening telomeres and does not contemplate measuring telomere length before and after administration of the supplement.
U.S. Pat. No. 7,955,626 to Pero (issued Jun. 7, 2011) discloses non-topical administration of a purified quinic acid alkyl ester or a carboxy alkyl ester. The reference discloses a water extract of Uncaria tomentosa, as well as further purification of the extract by thin layer chromatography to obtain carboxy alkyl esters (CAE). The invention is not directed to administration of a composition comprising quinic acid. Moreover, the method of the invention does not require the inclusion of quinic acid (in the free acid form or a salt thereof) because the invention is specifically directed to administering a pharmaceutical composition comprising an effective amount of an ester, either a quinic acid alkyl ester or a carboxy alkyl ester, meeting specified criteria. Furthermore, the reference does not disclose measuring telomere length and does not contemplate increasing the length of the telomeres.
U.S. Pat. No. 8,372,449 (Pero, Ronald W., issued Feb. 12, 2013) describes a method of non-topical administration of a pharmaceutical composition comprising a purified and isolated compound selected from the group consisting of quinic acid and a quinic acid salt. The invention is directed to a method for enhancing the response to tumor formation and/or growth in a mammal in need thereof. The pharmaceutical composition is administered in an amount effective to inhibit TNF-α production or to induce apoptosis of white blood cells, where said quinic acid and said quinic acid salt may form an ammonium salt and/or chelate thereof having a bioassay efficacy using IC50 in HL-60 cells of approximately 500 μg/ml or less. The disclosure indicates that the bioactive agent of C-MED-100® (a water extract of Uncaria tomentosa) in vivo is not quinic acid lactone, but rather is quinic acid and its salts, including its ammonium salt. The in vivo working examples pertain to oral dosing studies in mice and rats, with administration by gavage in rats and via the drinking water in mice. Treatment of mice with C-MED-100® resulted in an increased number of spleen cells, due to the prolongation of lymphocyte half life. In rats, quinic acid (QA) and its ammonium salt (QAA) were about as effective as C-MED-100® at reversing doxorubicin-induced leukopenia. The reference reveals that daily oral doses of C-MED-100® between 250-700 mg have proven efficacious in humans with regard to enhancing anti-inflammatory, DNA repair, immune stimulation, and anti-tumor processes. This reference does not contemplate a method of lengthening the telomeres or otherwise manipulating or maintaining telomere length. Likewise, the reference does not contemplate measuring telomere length before or after administration of quinic acid, a quinic acid salt, or C-MED-100®.
The bioactive component in AC-11®, formerly known as C-MED-100®, is reported to be quinic acid lactone (U.S. Pat. No. 6,964,784, issued Nov. 15, 2005). Moreover, the bioactive agent in vitro is quinic acid lactone (U.S. Pat. No. 8,372,448, issued Feb. 12, 2013), while the in vivo bioactive agent is quinic acid (U.S. Pat. No. 8,372,449, issued Feb. 12, 2013).
U.S. Pat. No. 8,974,839 to Giampapa (issued Oct. 16, 2014) discloses a multi-component supplement composition comprising a water extract of Uncaria tomentosa, which is disclosed as being a DNA repair complex. The supplement of the invention also comprises a telomere maintenance complex. However, the water extract of Uncaria tomentosa is not disclosed as being a telomere maintenance complex. The reference does not contemplate a method of lengthening the telomeres or otherwise manipulating telomere length. Likewise, the reference does not contemplate measuring telomere length.
U.S. Patent Application No. 2017/0128355 to Giampapa et al. (published May 11, 2017) discloses a method of topically applying a multi-component composition comprising a water soluble extract of an Uncaria species. The reference does not contemplate a method of lengthening the telomeres or otherwise manipulating or maintaining telomere length. Likewise, the reference does not contemplate measuring telomere length.
U.S. Pat. No. 5,656,665 to Yu et al. (issued Aug. 12, 1997) discloses a method for visibly reducing a skin wrinkle and for reversing the effect of aging on human facial skin by topically applying to the wrinkle and/or affected facial skin quinic acid or quinolactone. A wide variety of preparations are contemplated. The working examples are directed to testing the various compositions in 90 human volunteer subjects, by topical application to the skin. This reference does not contemplate a method of lengthening the telomeres or otherwise manipulating or maintaining telomere length. Likewise, the reference does not contemplate measuring telomere length before or after administration of quinic acid or quinolactone. The reference does not disclose administering a water extract of Uncaria tomentosa to a subject or a cell. Moreover, there is no mention of an extract of Uncaria tomentosa. The reference is limited to topical application and does not contemplate other modes of administration.
U.S. Patent Application Publication No. 2015/0338387 to Ehrenkranz (published Nov. 26, 2015) discloses a method for monitoring and adjusting a biometric, such as aging. The method of the invention comprises administering to a subject a supplement selected to effect the state of the selected biometric. The supplement may be formulated for oral administration. Among a long list of possibilities, the supplement may be an extract of Uncaria tomentosa and the biometric analyte, among a long list of possibilities, may be telomere length. The reference does not include any working examples, nor does it specifically mention using an extract of Uncaria tomentosa to increase the length of a telomere in a cell by administration of the extract to a mammal. Moreover, the reference does not specifically disclose a water extract of Uncaria tomentosa. 
U.S. Patent Application No. 2014/0079836 to McDaniel (published Mar. 20, 2014) discloses a method for modulating the activity of the gene maintenance process in order to influence the length and/or structural integrity of the telomere in living cells. Quinic acid is disclosed as an exemplary lifespan altering compound. The method of the invention is specifically directed to contacting a cell, tissue, organ, or organism with quinic acid, or an analog or derivative thereof. Quinic acid is disclosed as being naturally found in the coffee cherry. The disclosure contemplates lengthening telomeres, as well as modulating the activity or level of at least one of the telomere length maintenance genes or modulating the activity or level of telomerase. This reference does not contemplate using a water extract of Uncaria tomentosa for any purpose. There is no mention of an Uncaria species. The reference does not specifically point to quinic acid as modulating the length of the telomere in living cells. There are no working examples pertaining to quinic acid or telomere lengthening in an organism. Working examples directed to telomere length maintenance are provided, but these are in vitro experiments (cells in culture) examining the effects of green tea extract, coffee cherry extract, and idebenone.
It has now been shown by Applicants that administration of an aqueous extract of an Uncaria plant to a subject results in telomere lengthening.